基于肥大细胞-Th1/Th2/Th17细胞因子交互效应探讨中药防治银屑病瘙痒的分子协同机制及活性成分筛选

Itching is the most common complaint symptom in psoriatic patients besides lesions. Psoriasis is an immune disease mediated by T cells. Mast cells are important in innate immune response. The cytokines released by mast cells are significant factors causing itching, which can promote the secretion of cytokines by T cells and other white blood cells. In recent years, our research group has carried out clinical and basic research in vivo and in vitro on the basis of the famous doctor's prescription, Yin-Xie-Ling formula, and we found that it has effects of inhibiting keratinocyte proliferation, anti-inflammatory and anti-pruritic. In this study, for treating psoriasis pruritus, it was proposed a hypothesis “Yin-Xie-Ling formula regulating mast cells and Th1/Th2/Th17 cytokines might be the pharmacological mechanism to reduce psoriatic pruritus”. To verify this hypothesis, we employ network pharmacology to screen the key components of psoriatic components, and use molecular interaction devices to quantitatively determine the components of the screening components. At last,the potential active drugs will be evaluated separately in mice and mast cell models. The complement of the project will provide a more theoretical basis for explaining the material basis and regulatory mechanisms of traditional chinese medicine in treating psoriatic pruritus, and also important for Yin-Xie-Ling formula development and antipruritic drugs discovery.

瘙痒是银屑病患者除皮疹外最常见的主诉症状。银屑病是T细胞介导的炎症免疫性疾病，肥大细胞是参与机体固有免疫应答的重要细胞，肥大细胞脱颗粒释放的细胞因子是致痒的重要因素，可促进T细胞及其它白细胞分泌细胞因子形成瘙痒循环。近年课题组围绕名医验方银屑灵复方开展体内外的临床和基础研究，发现其具有抑制角质细胞增殖、抗炎、止痒作用。本研究从中药防治银屑病瘙痒的视角出发，提出“银屑灵复方调控肥大细胞与Th1/Th2/Th17细胞因子交互效应可能是其改善银屑病性瘙痒的药效作用机制”这一假说。为验证其科学性，本项目运用网络药理学方法对复方成分与银屑病关键靶点筛选，再结合分子相互作用仪对筛选成分进行定量，将得到的潜在活性药物分别在小鼠和细胞模型上评价。课题的完成将为分子水平阐释中药防治银屑病瘙痒的物质基础及其调控机制提供更完善的理论依据，对验方的开发、推广及止痒药物的发现均有重要的意义。

银屑病患者多伴有皮肤瘙痒，缓解瘙痒对提高银屑病患者的生活质量具有显著的意义。项目研究内容围绕验方银屑灵展开，运用网络药理学方法对复方成分与银屑病瘙痒关键靶点进行快速筛选，结合生物相互作用仪对筛选成分测定亲和力，筛选潜在活性药物于肥大细胞、动物模型评价。.本研究运用UPLC-MS技术明确了银屑灵复方102个天然分子化合物结构，并快速筛选小分子化合物与银屑病及瘙痒关键靶点(TNF-α,MAPK3,Inos,eNOS,mTOR,COX-2,Histamine H1 receptor,JAK,mTOR)作用的潜在活性成分和多靶点药物，将初步筛选的小分子成分结合炎症、增殖细胞模型结果进一步评价，11个小分子在分子相互作用仪与Th1/Th2/Th17代表细胞因子偶联测定亲和力，确定单体芍药内酯苷为候选止痒药物。细胞实验显示中药及候选药物不影响体外活化的肥大细胞释放组胺和P物质。动物实验结果：银屑灵高剂量YXL-H组对抑制急性瘙痒小鼠的搔抓反应与阳性西药MTX组相当，提示对急性瘙痒有缓解作用，且优于中剂量组，抗组胺药组优于阳性中成药组。小鼠组织病理：YXL-H组真皮血管增生明显减少。中剂量组、阳性中成药组以及抗组胺组较模型组改变不明显。根据研究结果提示组胺不是银屑灵复方及组分化合物改善银屑病瘙痒的主要作用靶点，肥大细胞释放的与Th相关的炎症、免疫调节因子可能是中药复方及其来源药物减轻瘙痒的核心。后续研究将围绕这个方向进一步展开。在项目的资助下，课题组开发了银屑病和皮肤瘙痒软件PsNPD v2.0、TIPP v1.0，均为非商业用途，可用于银屑病和皮肤瘙痒性疾病的的中药或复方的分子机制及开发等研究。